Organizer: Elizabeth Wright, Emory University
Lead Instructor: Kent McDonald
In this course, we will review why cryo-techniques for biological specimen preparation are superior to conventional methods. We will discuss high-pressure freezing, freeze substitution, and preserving fluorescence in polymerized resiins. Persons taking this course should leave with a better understanding of biological EM cryotechniques and their role in different applications such as correlative LM - EM, EM tomography, EM immunolabeling, and as the best method for preservation of cellular fine structure. Specimen preparation procedures for resin-based 3-D imaging methods will also be discussed.
Lead instructor: Caroline Miller
The requirements for successful immunohistochemical and immunocytochemical labeling vary widely with different biological systems. The optimal techniques for light-microscope labeling often differ greatly from those needed for electron microscopy. The basics of immunolabeling at the light- and electron-microscope levels will be presented, illustrated with examples from several different biological systems. Some of the more complex methods and applications used in electron microscopy will be discussed in depth. The course will cover specimen preparation, immunogold labeling and enhancement methods, multiple labeling and correlative LM/EM techniques.
Instructor: James Grande
This workshop covers a wide range of practical topics in the field of image analysis. Subjects will be covered in an easy-to-understand format so that users with little or no experience can understand how image analysis can provide extensive quantitative measurements that may lead to better understanding of material performance. Topics range from input devices to image-processing algorithms and how best to extract quantitative data. Treating image analysis as a problem-solving tool along with discerning key metrics within a microstructure is discussed through several real-life examples. Comparisons using ImageJ/Fiji and a commercial image analysis product will be demonstrated.
Lead Instructor: Cindi Schwartz
This workshop will cover the use of SerialEM for data acquisition and IMOD for 3-D reconstruction and analysis. For SerialEM, topics include basic operation, low dose mode, STEM imaging, use of direct detector cameras, tilt series acquisition, montaging, and automatic acquisition from multiple areas. For IMOD, topics include reconstruction from single and dual-axis tilt series, alignment of slices or tomograms from serial sections, automated processing of multiple tilt series, and modeling and visualization. Cryo and room-temperature applications will be covered. Although the emphasis will be on biological samples, many points are applicable to materials science.
Lead Instructor: Nestor J. Zaluzec
Imaging and spectroscopy have long played pivotal roles in characterization of materials in both the biological and physical sciences. Of course, a signal must absolutely be detectable from an object in order to make any observations or measurements. For this course, we will assemble a cadre of researchers from both academia and industry, who will discuss the principles of various imaging and spectroscopic detectors, their limitations, and future prospects and technologies. Topics will include optical sensors and single-particle sensors for photons, x-rays, electrons and ions, as well as electromagnetic-field sensors, and the use of these sensors as imaging/spectroscopic detectors.
Lead Instructor: John Mansfield
Variable-pressure scanning EM (VPSEM) and environmental scanning EM (ESEM), while readily available, are not used as frequently as they should be. This course will compare the structure, operation, and special detectors of the two. The practicality of x-ray spectroscopy will be examined. The unique contrast mechanisms in these microscopes will be discussed. The use of the VPSEM as an in-situ platform for hot, cold and mechanical testing experiments will also be covered. Applications experts from instrument manufacturers will be invited to describe new developments. A summary presentation comparing the VPSEM with conventional SEM will conclude the course.